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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

123
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Related Experiment Video

Updated: Aug 15, 2025

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
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Editing Aspergillus terreus using the CRISPR-Cas9 system.

Sra-Yh Shih1, Uffe Hasbro Mortensen2, Fang-Rong Chang1,3

  • 1Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung City, Taiwan.

Synthetic Biology (Oxford, England)
|December 30, 2022
PubMed
Summary

This study demonstrates CRISPR-Cas9 genome editing in Aspergillus terreus, enabling precise gene disruption, insertion, and deletion. This powerful tool facilitates gene function studies and molecular breeding in this important fungal species.

Keywords:
Aspergillus terreusCRISPR-Cas9genome-editing technologymarker-free donor DNA

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Area of Science:

  • Molecular Biology
  • Mycology
  • Biotechnology

Background:

  • CRISPR-Cas9 is a versatile tool for targeted mutagenesis in various organisms.
  • Its application in Aspergillus terreus for genome editing remains underexplored, lacking established guidelines.
  • Developing efficient genome editing protocols is crucial for functional genomics in A. terreus.

Purpose of the Study:

  • To establish and optimize CRISPR-Cas9 genome editing in Aspergillus terreus.
  • To demonstrate precise gene disruption, insertion, and deletion using the CRISPR-Cas9 system.
  • To provide a robust method for gene manipulation and functional studies in A. terreus.

Main Methods:

  • Construction of a modified single-guide RNA (sgRNA)/Cas9 expression plasmid for optimized gRNA expression.
  • Co-transformation of sgRNA/Cas9 plasmid with marker-free donor DNA for gene disruption and insertion.
  • Co-delivery of two sgRNA/Cas9 plasmids for gene deletion and DNA removal between targeting sites.

Main Results:

  • Precise disruption of lovB and lovR genes was achieved.
  • Targeted insertion of the lovF gene and iterative gene editing of lovF and lovR were successful.
  • Efficient deletion of ku70 and pyrG genes, and removal of DNA between targeting sites in pyrG were accomplished.

Conclusions:

  • The CRISPR-Cas9 system is a powerful and precise tool for genome editing in Aspergillus terreus.
  • The developed approach offers significant potential for targeted gene manipulation.
  • This methodology will greatly contribute to the functional study of genes in A. terreus.